LOCUS       sequence-449738-        5512 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1123m is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in pan-neuronal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5512)
  AUTHORS   Ben-Simon Y, Hooper M, Narayan S, Daigle TL, Dwivedi D, Way SW, 
            Oster A, Stafford DA, Mich JK, Taormina MJ, Martinez RA, Opitz-Araya
            X, Roth JR, Alexander JR, Allen S, Amster A, Arbuckle J, Ayala A, 
            Baker PM, Bakken TE, Barcelli T, Barta S, Bendrick J, Bertagnolli D,
            Bielstein C, Bishwakarma P, Bowlus J, Boyer G, Brouner K, Casian B, 
            Casper T, Chakka AB, Chakrabarty R, Chance RK, Chavan S, Clark M, 
            Colbert K, Collman F, Daniel S, Departee M, DiValentin P, Donadio N,
            Dotson N, Egdorf T, Fliss T, Gabitto M, Garcia J, Gary A, Gasperini 
            M, Gloe J, Goldy J, Gore BB, Graybuck L, Greisman N, Haeseleer F, 
            Halterman C, Haradon Z, Hastings SD, Helback O, Ho W, Hockemeyer D, 
            Huang C, Huff S, Hunker A, Johansen N, Jones D, Juneau Z, Kalmbach 
            B, Kannan M, Khem S, Kussick E, Kutsal R, Larsen R, Lee C, Lee AY, 
            Leibly M, Lenz GH, Li S, Liang E, Lusk N, Madigan Z, Malloy J, 
            Malone J, McCue R, Melchor J, Mollenkopf T, Moosman S, Morin E, 
            Newman D, Ng L, Ngo K, Omstead V, Otto S, Oyama A, Pena N, Pham T, 
            Phillips E, Pom CA, Potekhina L, Ransford S, Ray PL, Rette D, 
            Reynoldson C, Rimorin C, Rocha D, Ruiz A, Sanchez REA, Sawyer L, 
            Sedeno-Cortes A, Sevigny JP, Shapovalova N, Shepard N, Shulga L, 
            Sigler AR, Siverts L, Soliman S, Somasundaram S, Staats B, Stewart 
            K, Szelenyi E, Tieu M, Trader C, Tran A, van Velthoven CTJ, Walker 
            M, Wang Y, Weed N, Wirthlin M, Wood T, Wynalda B, Yao Z, Zhou T, 
            Ariza J, Dee N, Reding M, Ronellenfitch K, Mufti S, Sunkin SM, Smith
            KA, Esposito L, Waters J, Thyagarajan B, Yao S, Lein ES, Zeng H, 
            Levi BP, Ngai J, Ting JT, Tasic B
  TITLE     A suite of enhancer AAVs and transgenic mouse lines for genetic 
            access to cortical cell types.
  JOURNAL   Cell. 2025 May 16:S0092-8674(25)00513-6. doi: 
            10.1016/j.cell.2025.05.002.
  PUBMED    40403729
REFERENCE   2  (bases 1 to 5512)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2025 
            May 16:S0092-8674(25)00513-6. doi: 10.1016/j.cell.2025.05.002."
COMMENT     Sequence Label: AiP14292 -
            pAAV-AiE1123m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5512
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      865..874
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(871..873,874..876,877..1587)
                     /codon_start=1
                     /product="monomeric optimized variant of EYFP, identical to
                     mVenus but with L68V (Kremers et al., 2006)"
                     /label=SYFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1177..1196)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1524..1545
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1597..1653
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1666..1731
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1762..2139
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2226..2246)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2433..2544
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2662..2802
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2662..2791
                     /label=AAV2 ITR
     misc_feature    2809..2836
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    2837..2864
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     rep_origin      2933..3388
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3020..3039)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3230..3251
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3405..3424)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3524..3546
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3584..3602)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3670..3774
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3775..3843,3844..4635)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3993..4012)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4806..5394
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5295..5314
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5451..5478
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    5479..5506
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtatagtc acctcggggc cgccaagagc agccgctgcc
      181 gccctgccca cttccggcaa gccgcgggct cgagccgcca gggggcgcgc gcccaagctc
      241 aggggacaaa ggaagcgcag accggccgca ttattaccat aaaaggcaaa cgctggcccg
      301 gaggcggacc tagggcagga agccagacct ccgccccctc cccaaacggg cgcaagctcc
      361 gcctacactg cgccgacggg cggcttgtgc gcgcgcggga ggcgaccaca cccagcagga
      421 agcgcaaaca aggcgcccgt gccactgtga ccccgcccct tccggacaac ggccccgccc
      481 tcgcccaacc ccgggcagcc gcgctcccag cctcaatacg cacgcgcagc taactaggaa
      541 gagggggaga ggaagagctc cctccctagc gcacacgcgc ctcgatgccc agtgatagag
      601 agcgcacgcg cagtggcatc ccagccccca cccggccgag ccggccctgc cgcgcccgcc
      661 cgcccgcccg cccgcccggc aagccgaata ggcaaaccgg tttggacaaa gacccagagg
      721 ccattgaggc gtgatcgtag cgtctggttc ccaatactgt gtactctcaa ggagctcggg
      781 ctgggcataa aagtcagggc agagccatct attgcttaca tttgcttctg ggatccagat
      841 ctttcgaagc tagcgctacc ggtcgccacc atggtctcca agggcgagga gctgttcacc
      901 ggggtggtgc ccatcctggt cgagctggac ggcgatgtca acggccacaa gttcagcgtg
      961 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct gatctgcacc
     1021 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta cggcgtgcag
     1081 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     1141 gaaggctacg tccaggagcg caccatcttc ttcaaagacg acggcaacta caagacccgc
     1201 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     1261 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     1321 gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac
     1381 aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     1441 gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccaa gctgagcaaa
     1501 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     1561 actctcggca tggacgagct gtacaagggc tctggtgcta ccaacttctc actgttgaaa
     1621 caggcagggg atgtagagga gaatccaggg cctggtgcta gtggagacta caaagaccat
     1681 gacggagatt ataaagatca tgacatcgat tacaaggatg acgatgacaa gtccggactc
     1741 agatctggag gctccggagg cccagagcca gcgaagtctg ctcccgcccc gaaaaagggc
     1801 tccaagaagg cggtgactaa ggcgcagaag aaaggcggca agaagcgcaa gcgcagccgc
     1861 aaggagagct attccatcta tgtgtataag gttctgaagc aggtccaccc tgacaccggc
     1921 atttcgtcca aggccatggg catcatgaac tcgtttgtga acgacatttt cgagcgcatc
     1981 gcaggtgagg cttcccgcct ggcgcattac aacaagcgct cgaccatcac ctccagggag
     2041 atccagacgg ccgtgcgcct gctgctgcct ggggagttgg ccaagcacgc cgtgtccgag
     2101 ggtactaagg ccatcaccaa gtacaccagc gctaagtaat gaggcgcgcc gcggccgcga
     2161 attcgatatc ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     2221 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     2281 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt agttcttgcc
     2341 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     2401 actgacaatt ccgtggctcg agagatcttc gactgtgcct tctagttgcc agccatctgt
     2461 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     2521 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     2581 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgagatctc
     2641 acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     2701 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     2761 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgccta tcagcacaca
     2821 attgcccatt atacgcgcgt ataatggact attgtgtgct gatatgatgc ggtattttct
     2881 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     2941 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3001 gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3061 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3121 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     3181 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3241 ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt
     3301 ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3361 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     3421 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     3481 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     3541 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     3601 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     3661 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3721 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     3781 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     3841 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     3901 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     3961 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     4021 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     4081 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     4141 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     4201 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     4261 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     4321 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     4381 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     4441 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     4501 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     4561 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     4621 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     4681 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4741 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4801 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4861 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4921 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     4981 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5041 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5101 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5161 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5221 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5281 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5341 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5401 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca tatcagcaca
     5461 caattgccca ttatacgcgc gtataatgga ctattgtgtg ctgataacat gt
//
